Intro References Flashcards
Hoyle, Immunology, 2020
Took organotypic hippocampal brain slices in 3D and stained for microglia (IB4) and ASC-Citrine.
Can see when the NLRP3 inflammasome is activated, overtime the specks start to appear
Can also see the involvement of microglial processes which become more amoeboid
Lopez-Castejon and Brough 2011
During infections, pathogens can disrupt the ER-Golgi network, hindering the conventional secretion pathway. IL-1β, lacking a signal peptide, is synthesized in the cytosol and can be secreted via alternative routes that remain functional under such stress conditions
3 Models of IL-1B secretion
Cytokine Growth Factor Review, 2011
J Cell Sci, 2007
Used a Western block
- Lysate contained in the cell can see the precursor and then when is cleaved, leaves this
- Appears in the supernatant
- If genetically take the sequence in coding the mature IL-1, it just gets secreted, therefore there is something in the pro-domain that anchors it inside the cell.
Cell Death and Differentiation, 2016
- Transduce macrophages with lenti virus to express IL-1B (GFP)
- Also imaged dye propidium iodine which is added to cells that cannot enter the cell (only when the membrane is compromised)- becomes fluorescent when it binds to the DNA in the nucleus.
- Use ATP as a DAMP, as soon as the venous fluorescence disappears, the nucleus becomes fluorescent
- Can see in real time, lots of venous fluorescent means lots of pro-IL-1, but when activate, rapidly a pore forms and this pro-IL-1B leaves
- At this exact same time, the propidium iodine binds to the DNA and there is fluorescence here.
- Shows that IL-1 is secreted through a pore under these conditions.
Shi, 2015
Cleavage of GSDMD by inflammatory caspases determines pyroptotic cell death
iScience, 2023
Super resolution imaging of the inflammasome using nanobodies and STORM
Shows that there is a dense core of NLRP3 and a less dense periphery, know that ASC is similar.
Shows that we do not yet completely understand the interaction between the two as thought that a couple of NLRP3 molecules oligomerise the complete prion-like adaptor ASC
Parker et al 2002
The effect of treatment on the expression of downstream inflammatory mediators when applying IL-1
Can see there is a concentration dependent secreition of IL-1
If boil the preparation, inactivate the bioactive property of te molecule, there is no longer any increase, same if use an antagonist
Thornton et al, 2006
If culture neurons with IL-1 and look at the effect alone, they are not neurotoxic
If culture with pure astrocytes, they are not neurotoxic either
When cultured together, it induces a potent neurotoxic response which is concentration dependent on the effect of neuronal death
Desson et al 2003
IL-1 treatment on cultures of neurons with different concentrations increases the rate fo neuronal firing
Due to opening of the NMDA channel
Wong et al 2019
Endothelial cell specific IL-1R1 deletion with tamoxifen
target the exome of the IL-1R1 gene and flank with a LOXP site
Cross this with a creorecombinase mouse in for brain endothelial cells only
Get a mouse lacking in endothelial cells only when induce using tamoxifen
Then do an experimental injury such as a stroke, reduce the brain damage as seen by reduced igG staining (marker for BBB dysfunction)
Decreased expression of ICAM and the block of neutrophil migration into the brain too
